
Description:
KAPAStrandedmRNA-SeqKit,withKAPAmRNACaptureBeadsKAPAStrandedmRNA-SeqKits
Evendifficultmessagesshouldbeunderstood.
KAPAStrandedmRNA-SeqKitsincludealltheenzymesandbuffersrequiredforCDNAlibrarypreparationforIlluminaNext-GenerationSequencing,utilizing100ng–4µgoftotalRNA.KAPAmRNACaptureBeadsareincludedforisolationofpoly(A)-tailedRNA.Kitsprovideprecisemeasurementofstrandorientation(>99%),uniformcoverage,andhigh-confidencemappingofalternatetranscripts,andareoptimizedfortheimprovedcoverageofGC-richandlow-abundancetranscripts.*KitscontainKAPAHiFiforhigh-efficiencyandlow-biaslibraryamplification,aswellasKAPAmRNACaptureBeadsandastreamlined,“with-bead”protocol.
KAPAStrandedRNA-SeqKitsincludealltheenzymesandbuffersrequiredforcDNAlibrarypreparationforIlluminaNext-GenerationSequencing,butdonotcontaintheKAPAmRNACaptureBeads.Kitscanbeusedtopreparelibrariesfrom10–400ngofeitherpoly(A)-selected,ribosomallydepleted,ortotalRNA.
NEW!KAPADual-IndexedAdapterKitsarenowavailable.FormoreinformationonKAPAAdapterKits,scrolldowntotheOrderingsection,ordownloadtheKAPAAdapterandBeadCalculator.
Download AdapterandBeadCalculator
*Dataonfile.
ForResearchUseOnly.Notforuseindiagnosticprocedures.
ProductHighlights

Uncoverchallengingtranscripts
- ImprovedcoverageofGC-richtranscripts
- Enhancedidentificationofexonicregions

Detectlow-abundancetranscripts
- Enablesidentificationoftranscriptsmissedbycompetitorkits,evenwithhighinput*
- Highuniformityacrossvaryingamountsofsampleinput*

Identifymoregenes
- HigherpercentageofuniquelymappedreadscomparedtoIllumina TruSeq™StrandedmRNASamplePrepKits*
- Lowerduplicationratesyieldbettercoverage

Maintainhighcoverageuniformity
- Minimal5’–3′biasacrosstranscripts
- Moreuniformdistributionofreadsovereachtranscript
*Dataonfile.
RelatedProducts
SequencingchallengingRNAsamples?HowaboutDNA?CheckouttheseKapaNGSproductstoimproveyourworkflowandresults:

KAPAhgDNAQuantificationandQCKit

KAPALibraryAmplificationKits

KAPAHyperPrepKits

KAPALibraryQuantificationKits
Applications:
Applications- Geneexpression
- Singlenucleotidevariation(SNV)discovery
- Post-transcriptionalSNVs
- Fusiongeneidentification
- Targetedtranscriptome
- Wholetranscriptome
KitSpecificationsandContents/Storage:
KitSpecificationsandContents/StorageKitscanbestoredforupto8 months at-20˚C. KAPAmRNACaptureBeadscanbestoredforupto8months at-4˚C.
Components
Specifications
- SpecDescription
- CompatIBLePlatformIlluminaHiSeq,NextSeq,MiSeqandGAIIx
- StartingMaterialTotalRNA,mRNA,orribosomaldepletedRNA
- InputAmountStrandedmRNA-SeqKits:100ng–4µgStrandedRNA-SeqKits:10ng–400ng
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1、测浓度和纯度,是否达标
2、跑电泳看条带是否与测的数据相互印证
如果上述两点满足,就不需要纯化,否则需要进一步纯化。
不过根据经验,一般都不需要,我们实验室用的是GNT系列的试剂盒,提取结果就直接进入下游实验了,效果挺稳定
质粒是真核细胞细胞核外或原核生物拟核区外能够进行自主复制的遗传单位,包括真核生物的细胞器(主要指线粒体和叶绿体)中和细菌细胞拟核区以外的环状脱氧核糖核酸(DNA)分子.现在习惯上用来专指细菌(大肠杆菌)、酵母菌和放线菌等生物中细胞核或拟核中的DNA以外的DNA分子.
小鼠没有叶绿体,你可以直接参照小鼠线粒体的提取方法进行提取.
你可以用你的目的引物,用反转录第一链进行扩增,看看能不能得到目的片段.
提RNA一般注意:提取前材料的保存(新鲜材料,过夜处理的样品),提取中对外源性的RNA酶的清除控制(离心管,PCR管,电泳槽,台面等的无RNA酶化处理),提取后需要-80度保存.其次跑电泳时,注意电泳液最好用新鲜的,核酸染料等诸多因素.
细菌总RNA提取试剂盒( germs islation kit).
对于外源性RNA酶的控制外源RNA酶清除剂,避免使用致癌物DEPC的危险方法就可简单操作清除耗材,台面,仪器等的外源RNA酶的降解作用.
向左转|向右转
目的掌握RNA提取的基本技术,了解RNA提取过程中的各种注意事项。二、原理RNA提取是分子生物学实验中难度较大的实验技术。RNA提取和DNA提取有类似的地方,因为它们都是核酸,都具有较好的水溶性。提取RNA首先破碎细胞,然后用提取液将RNA溶出,反复抽提去除蛋白质 ……
人教版高中生物必修二有这个图
实验非常不顺,想构建CDNA文库,但是从mRNA开始屡次失败,考虑主要是纯化过程中损失过多。想从总RNA入手,但是不知道实验步骤。不知那位大侠能提供总RNA建库的实验步骤。另外我现在手头有OligoDT,RT酶,苦于没有第二连合成试剂盒,不知道能否用普通PCR试剂盒Teq酶替代第二链合成过程中的DNA聚合酶I。好像有种方法合成第二链时,不需要另外的引物,利用降解的RNA作引物即可,我想直接设定PCR两个循环,合成第二链,不知方法可行否?愁啊,等着毕业,时间紧急,恳请帮忙。谢谢。

